Hepatocellular carcinoma (HCC) is the most common malignancy of the liver, which can progress rapidly and has a poor prognosis. Glypican-3 (GPC3) has been proposed to be an important diagnostic biomarker and therapeutic target for HCC. Aptamers have emerged as promising drug delivery vehicles because of their high binding affinity for target molecules. Herein, we developed G12msi, a gemcitabine-incorporated DNA aptamer, targeting GPC3, and evaluated its binding specificity and anti-tumor efficacy in GPC3-overexpressing HCC cell lines and murine xenograft models. GPC3-targeted aptamers were selected by using the SELEX process and the chemotherapy drug gemcitabine was internally incorporated into the aptamer. To determine the binding affinity and internalization of the G12msi, flow cytometry and confocal microscopy were performed on GPC3-positive HepG2, Hep3B, and Huh7 cells, as well as a GPC3-negative A431 cell. The anti-tumor activities of G12msi were evaluated with in vitro and in vivo models. We found that G12msi binds to GPC3-overexpressing HCC tumor cells with high specificity and is effectively internalized. Moreover, G12msi treatment inhibited the cell proliferation of GPC3-positive HCC cell lines with minimal cytotoxicity in control A431 cells. In vivo systemic administration of G12msi significantly inhibited tumor growth of HCC HepG2 cells in xenograft models without causing toxicity. These results suggest that gemcitabine-incorporated GPC3 aptamer-based drug delivery may be a promising strategy for the treatment of HCC.

中文翻译：

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吉西他滨合并GPC3适体的靶向治疗肝癌

肝细胞癌（HCC）是最常见的肝脏恶性肿瘤，可以快速进展并且预后较差。Glypican-3（GPC3）已被提出是肝癌的重要诊断生物标志物和治疗靶标。由于适体对靶分子的高结合亲和力，它们已成为有前途的药物递送载体。在这里，我们开发了靶向GPC3的吉西他滨结合的DNA适体G12msi，并在过表达GPC3的HCC细胞系和鼠异种移植模型中评估了其结合特异性和抗肿瘤功效。通过使用SELEX方法选择靶向GPC3的适体，将化疗药物吉西他滨内部掺入适体中。要确定G12msi的结合亲和力和内在化作用，在GPC3阳性的HepG2，Hep3B和Huh7细胞以及GPC3阴性的A431细胞上进行了流式细胞术和共聚焦显微镜检查。用体外和体内模型评估了G12msi的抗肿瘤活性。我们发现，G12msi以高特异性结合过表达GPC3的HCC肿瘤细胞并被有效地内化。此外，G12msi处理可抑制GPC3阳性HCC细胞系的细胞增殖，且对对照A431细胞的细胞毒性极小。在异种移植模型中，体内全身施用G12msi可显着抑制HCC HepG2细胞的肿瘤生长，而不会引起毒性。这些结果表明，结合吉西他滨的基于GPC3适配子的药物递送可能是治疗HCC的有前途的策略。用体外和体内模型评估了G12msi的抗肿瘤活性。我们发现，G12msi以高特异性结合过表达GPC3的HCC肿瘤细胞并被有效地内化。此外，G12msi处理可抑制GPC3阳性HCC细胞系的细胞增殖，且对对照A431细胞的细胞毒性极小。在异种移植模型中，体内全身施用G12msi可显着抑制HCC HepG2细胞的肿瘤生长，而不会引起毒性。这些结果表明，结合吉西他滨的基于GPC3适配子的药物递送可能是治疗HCC的有前途的策略。用体外和体内模型评估了G12msi的抗肿瘤活性。我们发现，G12msi以高特异性结合过表达GPC3的HCC肿瘤细胞并被有效地内化。此外，G12msi处理可抑制GPC3阳性HCC细胞系的细胞增殖，且对对照A431细胞的细胞毒性极小。在异种移植模型中，体内全身施用G12msi可显着抑制HCC HepG2细胞的肿瘤生长，而不会引起毒性。这些结果表明，结合吉西他滨的基于GPC3适配子的药物递送可能是治疗HCC的有前途的策略。G12msi处理可抑制GPC3阳性HCC细胞系的细胞增殖，且对对照A431细胞的细胞毒性极小。在异种移植模型中，体内全身施用G12msi可显着抑制HCC HepG2细胞的肿瘤生长，而不会引起毒性。这些结果表明，结合吉西他滨的基于GPC3适配子的药物递送可能是治疗HCC的有前途的策略。G12msi处理可抑制GPC3阳性HCC细胞系的细胞增殖，且对对照A431细胞的细胞毒性极小。在异种移植模型中，体内全身施用G12msi可显着抑制HCC HepG2细胞的肿瘤生长，而不会引起毒性。这些结果表明，结合吉西他滨的基于GPC3适配子的药物递送可能是治疗HCC的有前途的策略。